Immunohistochemistry analysis of Nanog showing staining in the nucleus of paraffin-treated human testicular carcinoma (right) compared with a negative control in the absence of primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Nanog polyclonal antibody (ab155943) diluted by 3% BSA-PBS at a dilution of 1:500 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
All lanes : Anti-Nanog antibody (ab155943) at 1/2000 dilutionLane 1 : NCCIT whole cell lysateLane 2 : NTERA-2 whole cell lysateLane 3 : HeLa whole cell lysateLysates/proteins at 80 µg per lane.
Immunofluorescent analysis of either Human embryonal carcinoma NTERA-2 cells (right) or negative control HeLa cells(left), labeling Nanog with ab155943 1/200 dilution.
Immunofluorescent analysis Human embryonic stem cell line H9, labeling Nanog with ab155943 at 1/100 dilution (red). DAPI staining (pink).
Immunoflurorescent analysis of matrigel coated and formaldehyde fixed Human iPS cell line HEL 11.4, labeling Nanog with ab155943 at dilution 1/100 (red). DAPI staining (pink).